2010創世紀專刊_RNA Extraction_100708

RNA From RNA-Bee MasterPure Complete DNA and RNA Purification Kit NucleoSpin RNA II NucleoSpin RNA L

From LCM sample NucleoSpin RNA XS ArrayPure Nano-scale RNA Purification Kit

From MasterPure Yeast RNA Purification Kit

Small RNA Application NucleoSpin miRNA

From NucleoSpinRNA Plant MasterPure Plant RNA Purification Kit

From FFPE sample WaxFree RNA NucleoSpin FFPE RNA QuickExtract FFPE RNA Extraction Kit

QuickExtract FFPE RNA Extraction Kit QuickExtract RNA Extraction Kit

mRNA Isolation NucleoSpin mRNA MiniMidi mRNA-ONLY Prokaryotic/Eukaryotic mRNA Isolation Kit

RNA Clean-Up NucleoSpin RNA Clean-UpRNA Clean-UP XS

NucleoSpin Triprep( RNA/DNA/Protein) NucleoSpin RNA/Protein

RNARNA

BioBio--Genesis Technology Genesis Technology

P.1 - P.3

P.4 P.5 - P.6

P.7 - P.8 P.9

P.10 P.11

P.12

P.13 - P.14

P.15 - P.16 P.17

P.18 P.19 - P.20

P.21

P.21

P.22

P.23 P.24

P.25 - P.26

P.27

P.28 P.29

RNA

RNA RNA

-80oC RNA RNase

RNA RNA

RNase RNA RNA (RNA

extraction kit)

RNA Guanidinium thiocyanate RNase

phenol-chloroform (pH=4.5) RNA single reagent (ex: RNAzol Bee)

Single reagent RNA

single reagent -80oC< 1 month RNA RNA

RNA single reagent RNA

RNA genomic DNA phenolchloroform

RNA

RNA

RNA silica column RNA

RNA

Silica column silica

silica column RNA DNase I genomic DNA

wash buffer

RNASilica column RNA column silica

column RNA column nucleotide fragment size binding capacity

Epicentre MasterPure RNA extraction kit silica column

MasterPure lysis buffer proteinase K

isopropanol nucleotide RNase-Free DNase I

DNA total RNA silica column MasterPure

RNA

RNA RNA TE buffer TE buffer EDTA RNA

RNA DEPC-treated water nuclease-free water-

80oC RNA RNase

RNA RNA RNase inhibitor (ex: RiboGuard RNase in-

hibitor) RNA

RNA RNA RNA RNA

RNA RNA

RNA

1.

RNA 230nm260nm 280nm 260nm

DNA RNA280nm 230nm A260

RNA (RNA concentration = A260 x dilution fold x 40 ng/ul) RNA

A260/A280 ratio 1.9~2.1 A260/A230 ratio 2~2.4 A260/A280 1.9

sample

RNA RNA

A260 RNA

2.

RNA sample

genomic DNA rRNA RNA

RNA

RNA

RNase-Free RNase RNA

RNA single-strand nucleotide

RNA rRNA 0.5x TBE

agarose gel RNA size

formaldehyde denature agarose gel

RNA P.2P.2

3. RNA Integrity Number (RIN)

RNA Integrity Number RNA RNA

RNA RINRIN

1~10 RNA

RNA A. silica column solution method

RNA silica column

RNA solution method

B.

RNA BeeNucleoSpin RNA II or MasterPure RNA purification kit

NucleoSpin RNA XS or ArrayPure Nano-Scale RNA purification

kit

NucleoSpin Plant or MasterPure Plant purification kit

FFPE sample NucleoSpin FFPE or QuickExtract FFPE RNA extraction kit

C.

miRNA assay NucleoSpin miRNA

QuickExtract RNA extraction kit

RNA Clean-Up NucleoSpin RNA Clean-Up or RNA Clean-Up XS

mRNA isolation NucleoSpin mRNA Mini or mRNA Midi

Fig.1 RNA Integrity Number (RIN)

Total RNA RIN 10 RIN 2 RIN

10 sample 18S 28S rRNA

RIN rRNA RNA RIN

2 rRNA RNA

RNA

RIN Value Most Suitable Application

1 ~ 4 PCR assays with short region of amplification

4.1 ~ 6.9 qRT-PCR applications

7~10 Highly demanding gene array assays

RNA RIN

RNA P.3P.3

Traditional RNA Isolation

RNA-Bee single reagent RNA 1

/

1ml RNA-Bee 50 mg 5 x 106 100 l whole blood 109

MN NucleoSpin RNA Clean-up or NucleoTrap mRNA

Ordering Information

Tel-Test RNA-Bee 200ml LB-CS-105B

RNA-Bee easy protocol

RNA-Bee phenol quanidine thiocyanate

single reagent RNA-Bee

chloroform //

DNA

RNA isopropanol RNA

RNA pellet RNase-Free water

-70oC RNA reverse transcrip-

tionRT-PCRNorthern Blot

From P.4P.4

MasterPure Complete DNA and RNA Purification Kit

MasterPureTM Complete DNA and RNA purification kit total nuclei acid

(TNA)DNA RNAMasterPure

TNA DNase I total

RNA RNase A genomic DNA RNA gDNA kit

30min TNA60min total RNA column

phenolchloroform

(include

Fig.3 MasterPure sample(M= 1kb ladder)


MasterPure Complete DNA and RNA

Purification Kit

For 10 gDNA or 10 TNA or 5 RNA purifications MC89010

For 200 gDNA or 200 TNA or 100 RNA purifications MC85200

MasterPure RNA Purification Kit 100 RNA purifications MCR85102

MasterPure DNA Purification Kit

(for isolating TNA or DNA) 200 gDNA or 200 TNA purifications MCD85201

Sample Sample Size TNA (g) DNA (g)

RNA (g)

HeLa/HL60 cells 1 X 106 cells 10-30 3-12 7-15

Tissues

Liver 5 mg 33-42 5-10 13-25

Brain 5 mg 9-13 6-9 4-11

Heart 5 mg 6-10 4-7 4-5

Kidney 5 mg 10-17 3-8 14-17

Thymus 5 mg 15-30 6-12 9-18

Blood 200 l 3-10 3-9

Buffy coat 300 l 40-55 40-55 3-6

Mouse tail 0.5 cm 25-30 9-11

E. coli 3.5 x 106 cells 2.5-2.8 1.3-1.6 1.6-1.8

S. mutans 1.5 ml 0.9

Yeast (S. cerevisiae)

2.2 x 106 cells 11-18

1.1 x 107 cells 70-78

Table.2 MasterPureTM Complete DNA and

RNA purification kit

Targets Samples Extracted

Human & mammalian RNA & genomic DNA HIV HCV RSV HPV Yeast Enterovirus B. pertussis E. coli M. tuberculosis Streptococcus mutans Soy (tofu) Maize Insect tissues

Plasma Whole blood Guthrie cards Buccal cells Liver Mouse tail Kidney Saliva Urine Sputum Tissue-culture cell lines Cervical cells Paraffin-embedded tis-sues

Table.1 MasterPureTM Complete DNA and RNA purifica-

tion kit

From P.6P.6

NucleoSpin RNA II NucleoSpin Filter RNase-Free DNase column genomic DNA

DNA

(G+/G bacteria) RNA

phenol/chloroform

30 total RNA

Fig.2 MN NucleoSpin RNA II Q

Total RNA 106 HeLa cells MN NucleoSpin RNA II (Lane 1-3) Q RNA kit

(Lane 4-6) elute 50ul 10ul 1.2% Formadelhyde gel

Fig.1 Total RNA

1.2ml LB cultured Pseudomonas putida (OD600=0.9)

0.05mg/ml (1&4), 0.2mg/ml (2&5) and 0.8mg/ml

(3&6) lysozyme 37oC 10min NucleoSpin RNA II

total RNA 6ul total RNA 1.2% formaldehyde gel

A260/280 2.1

From P.7P.7

Fig.3 ()MN NucleoSpin RNA II 10 cells total RNA RT-PCR

MN NucleoSpin RNA II 105, 104, 103, 330, 100, 33, 10, 3 0 HeLa cell

total RNA (Lane 1~9) 3ul total RNA b-actin primer RT-PCR 15ul 1.2%

agarose gel 10 HeLa cellsNucleoSpin RNA II RT-PCR

total RNA


NucleoSpin RNA II 10 reactions 740955.10

50 reactions 740955.50

250 reactions 740955.250

Fig.4 Genomic DNA Q

Total RNA 106 HeLa cells MN NucleoSpin

RNA II (Lane 1-3) Q RNA kit (Lane 4-6) 10ul

total RNA RNAse A 1% TAE agarose gel

genomic DNA Q Dnase I total RNA

samplegenomic DNA MN NucleoSpin

RNA II

Technology Silica membrane technology

Format Mini spin columns

Sample material < 5 x 106 cultured cells, < 109 bacterial cells

< 108 yeast cells, < 30 mg tissue

Fragment size > 200 b

Typical yield 14 g from 106 HeLa cells

70 g from 109 bacterial cells

A260/280 1.9-2.1

Typical RIN (RNA integrity number) > 9

Elution volume 40-100 l

Preparation time 30 min/6 preps

Binding capacity 200 g

Product Information

Fig.5 rDNase MDB (membrane desalting buffer) genomic

DNA

NucleoSpin RNA II total RNA MDB (+MDB)

MDB (-MDB) rDNase DNA contaminant assay

qPCR PCR product agarose gel sample

DNA PCR rDNase MDB

gDNA contamination

From P.8P.8

NucleoSpin RNA L 200mg total RNA

600ug RNA

phenol/chloroform

(G+/G bacteria)


Format Midi spin columns

Sample material < 5 x 107 cultured cells, < 200 mg tissue

< 1010 bacterial cells, up to 3 x 108 yeast cells

Fragment size 200b ~ 20Kb

Typical yield 180 g from 107 HeLa cells

620 g from 4 x 107 HeLa cells

A260/280 1.9-2.1

Typical RIN (RNA integrity number) > 9

Elution volume 500 l



Product Information

NucleoSpin RNA L 20 reactions 740962.20

Fig.1 NucleoSpin RNA L total RNA

Pig liver & kidney NucleoSpin RNA L Q

total RNA

A: RNA isolated from 100mg (1~4) and 200mg (5~8) pig

kidney.

Lane 1,2,5,6: NucleoSpin RNA L

Lane 3,4,7,8: Competitor Q

B: RNA isolated from 200mg pig liver sample.

Lane 1,2,3: NucleoSpin RNA L

Lane 4: Blank

Lane 5,6,7: Competitor Q

From P.9P.9


NucleoSpin RNA XS

0.1mg total

RNA


Format XS spin columns

Sample material Tissue 200b

Typical yield 102 HeLa cells: 0.1~1.5ng ; 103 HeLa cells: 10~15ng

104 HeLa cells: 100~150ng ; 105 HeLa cells: 1000~1500ng

A260/280 1.9-2.1

Typical RIN

(RNA integrity number) > 9 (Depending on sample quality)

Elution volume 5-20 l



Product Information

Total RNA Extraction

NucleoSpin RNA XS 10 reactions 740902.10


250 reactions 740902.250


Tissue or Cell

Source

Amountof Start-

ing Material

NucleoSpin RNA XS

SampleNo. 1 (ng/ l)

NucleoSpin RNA XS

SampleNo. 2 (ng/ l)

Competitor Q Sam-

pleNo. 1 (ng/ l)

Competitor Q Sam-

pleNo. 2 (ng/ l)

Mouse Kidney 0.5 mg 157 108 96 63

Mouse Kidney 0.1 mg 27 25 17 18

HeLa Cells 1,000 cells 1.3 1.7 0.08 0.06

Table.1 sample NucleoSpin RNA XS Q total RNA

ArrayPure Nano-scale RNA Purification Kit

ArrayPureTM Nano-scale RNA purification kit (1-104 cells) RNA pu-

rification kit RNA column

LCM sample total RNA real-time RT-PCR 2-round aRNA synthesis

for microarray analysis

Fig.1 ArrayPureTM Nano-scale RNA purification kit 102101 HeLa

cells total RNA(3)RNA MMLVcDNAreal-time

PCR

Fig.3 Fig.1 data standard curveSlope=-3.14R2=0.992

Fig.2 ArrayPureTM Nano-scale RNA purification kit

flask HeLa cell RNA label

Cy3Cy5 microarray analysis

0.946


10 RNA purifications MPR09010

100 RNA purifications MPR09100

: Nano-scale Lysis Solution, 2X Nano-scale Lysis Solution, MPC Protein Precipitation Reagent, Proteinase

K, RNase-Free DNase I, RiboGuard RNase Inhibitor, 1X DNase Buffer, TE Buffer.

ArrayPure Nano-scale RNA Purification Kit

From LCM Sample P.11P.11

MasterPure Yeast RNA Purification Kit

MasterPureTM Yeast RNA purification kit

RNA

phenol

RNA

1ml S.cerevisiae 25~50ug RNA

cDNA synthresismicroarray gene expression assay

Quality of obtained using the MasterPure Yeast RNA Purifica-

tion Kit.

MasterPure Yeast RNA Purification Kit S.cerevisiae

-20oC 2 Agilent Technologies 2100 Bioanalyer

MasterPure Yeast RNA Kit

RNA RNA

Microarray analysis of yeast RNA purified with the MasterPure Yeast

RNA Purification Kit.

S.cerevisiae MasterPure Yeast RNA Purification Kit RNA

reverse transcriptionlabeling hybridized to S. cerevisiae DNA mi-

croarray


10 RNA purifications MPY83010

100 RNA purifications MPY83100

: Extraction Reagent for RNA, MPC Protein Precipitation Reagent, TE Buffer (in 100-reaction kit only), Pro-

teinase K, RNase-Free DNase I, RiboGuard RNase Inhibitor, 10X DNase Buffer, 2X T & C Lysis Solution.

MasterPure Yeast RNA Purification Kit

From P.12P.12

NucleoSpin miRNA Small RNALarge RNA

RNA

Small RNA (18~200 bases)

Small RNA (18~200 bases) Large RNA(>200bases)

total RNA ( Small RNA Large RNA)

SDS-PAGE Western Blot

Phenol/Chloroform RNA

chaotropic salt

spin-column

NucleoSpin Filter:

NucleoSpin Protein Removal Columns: RNA

rDNase: column genomic DNA

Fig.2 Agilent Bioanalyzer MN NucleoSpin miRNA total RNASmall RNA Large RNA

107 HeLa cell NucleoSpin miRNA RNA total RNA (A)small RNA (B) arge RNA (C)NucleoSpin miRNA

total RNA small RNA large RNA

Fig.3

NucleoSpin

miRNA 104

105106107

HeLa cell total

RNA MiR-

16 qPCR

total RNA

qPCR

Ct RNA

Fig.4

NucleoSpin miRNA

Laemmli-like PSB

40ug 12% SDS-PAGE

1. mouse liver

2. mouse kidney

3. mouse spleen

4. mouse lung

5. mouse heart

6. porcine liver

7. human brain

8. HeLa cell

Fig.1 2.5ug, 5ug, 10ug, 15ug RNA DICER enzyme small RNA

NucleoSpin miRNA large dsRNA small siRNA 1%

TAE agarose gel

1,12:100bp DNA marker 2,11:unpurified DICER reaction mix

3-6: 25ul purified large dsRNA 7-10: 25ul purified small RNA

Small RNA Application P.13P.13

Small RNA only Small RNA and Large RNA Total RNA

miRNA



Sample material < 107 culture cells, < 30mg human/animal tissue

< 50mg plant tissue, < 150ul reaction mixture

Fragment size for Small RNA 18 - 200 bases

Fragment size for Large RNA >200 bases

Typical yield 10ug small RNA, 95ug large RNA from 107 HeLa cells

Binding capacity 200g

Elution volume 30 - 100 l

Preparation time < 45 min (6 preps human/animal tissue, small and large RNA)

< 35 min (6 preps human/animal tissue, small RNA)

Product Information


NucleoSpin miRNA 10 reactions 740971.10


250 reactions 740971.250

Fig.5 NucleoSpin miRNA gDNA Q A

107 HeLa cells NucleoSpin miRNA

Q A miRNA extraction

qPCR 200bp ATPase 6 gene

NucleoSpin miRNA gDNA Ct A (

Ct=3.5) Q ( Ct=4.3) A Q gDNA

NucleoSpin miRNA 10

MN Q A

Small RNA ApplicationP.14P.14

miRNA Extraction

NucleoSpin RNA Plant

Plant RNA Extraction

RNA

Total RNA

2 lysis buffer (RA1RAP)

70ug RNA

Real-Time RT-PCRprimer extensionarray technologyRNase protection as-

say

Lanes: Average Yield

1-3: poppy seed lysed with buffer RA1 0.84ug

4-6: poppy seed lysed with buffer RAP 7ug

Fig.4 NucleoSpin RNA Plant 2 lysis buffer

lysis buffer RNAMN NucleoSpin RNA Plant RA1

RAP lysis buffer

MucleoSpin RNA Plant 50mg RA1 (Lane 1~3) RAP (Lane 4~6) lysis

total RNA 20ul total RNA 1.2% Formaldehyde agarose gel RAP

lysis RA1 total RNA

Fig.1 NucleoSpin RNA Plant RT-PCR inhibitor

ex: RT-PCR

16 RT-PCR 8 50mg NucleoSpin

RNA Plant RNA 60ul elute 3ul total RNA RT

-PCR 15ul PCR product 1.2% agarose PCR product

200bp.

M: marker 1,2: Tobacco 3,4: Tomato 5,6: Red pepper 7,8: Zucchini

9,10: eggplant 11,12: Broccoli

13,14: Radish 15,16: Maize

Fig.2 NucleoSpin RNA Plant sample total RNA

50mg NucleoSpin RNA Plant total RNA

1% Formaldehyde gel

Lane 1: maize,

Lane 2: thyme,

Lane 3: Palma christi

Lane 4: tobacco,

Lane 5: rye,

M: Marker.

Fig.3 NucleoSpin RNA Plant Q

100mg NucleoSpin RNA Plant Q

total RNA 100ul 10ul 1.2% formaldehyde

agarose gel A260/280 1.8~2.1

NucleoSpin RNA Plant RNA Q

From P.15P.15



Sample material 1-100mg tissue

Fragment size 200 b - 20kb

Typical yield 3-70ug from 100mg plant material

A260/280 1.9-2.1

Elution volume 60 l



Product Information


NucleoSpin RNA Plant 10 reactions 740949.10


250 reactions 740949.250

NucleoSpin RNA Plant

Table.1 50mg total RNA

From P.16P.16

MasterPure Plant RNA Purification Kit

MasterPureTM Plant RNA purification kit Plant Tissue and Cell Lysis Solution

WaxFreeTM RNA xylene

column resin RNA RNA

2hr

FFPE samplefresh or frozen tissuecultured cells

RT-PCRReal-Time RT-PCR

WaxFreeTM RNA FFPE sample RNA extraction kit silica column

RNA RNA binding elute RNA

WaxFree RNA Q-solution paraffin enzyme mix R-resin

DNA RT-PCR total RNA

WaxFree RNA

WaxFree RNA extraction from lung tissue FFPE section

WaxFree RNA FFPE sample (size: 1x1 cm5um thickness)

total RNA 20ul 50ul RT RT 15ul 50ul PCR

primer EGFR primer setPCR 195bp

Lane 1:100bp ladder;

Lane 2: NTC

Lane 3: gDNA control; .

Lane 4: WaxFree RNA extract

Lane 5: WaxFree RNA extract without tissue

Lane 6: Commercial total RNA (positive control)


WaxFreeTM RNA 50 reactions WR-50

From FFPE Sample P.18P.18

-RNA Extraction from paraffin sample

1ul RT product 30ul qPCR reaction

Paraffin Dissolver paraffin xylene 3

paraffin

Decrosslinking bufferrDNase DNA

XS mini column 5ul RNA RNA

70

RT-PCRReal-Time RT-PCR

FFPE RNA DNA NucleoSpin FFPE RNA/DNA

NucleoSpin FFPE RNA

Table.1 Q R NucleoSpin FFPE RNA

RNA quality


NucleoSpin FFPE RNA 10 reactions 740969.10


250 reactions 740969.250


NucleoSpin FFPE RNA/DNA 10 reactions 740978.10

250 reactions 740978.250

Paraffin Disslover 25ml 740968.25



Format Mini spin columns - XS design

Sample material Up to 7 sections (10um)of 250mm2 or 15mg paraffin

Typical yield Depend on amount and quality of sample

A260/280 1.9 - 2.1

Typical RIN 2 - 6 (Strongly depend on sample quality)

Elution volume 5 - 30 l

Preparation time ~70 min/6 preps


Product Information


FFPE RNA FFPE RNA Extraction

7.5mm FFPE

RNA

NucleoSpin FFPE RNAQ

R kit MN

NucleoSpin FFPE RNA

RNA

RNA RNA Q

R

QuickExtract FFPE RNA Extraction Kit

32 FFPE sample RNA

xylenephenol

DNase I

RT-PCR

Fig.1 QuickExtractTM FFPE RNA

Extraction kit FFPE sample from

skeletal muscle RNA MMLV

cDNA PCR

primer set PCR

Lane M, 100-bp DNA ladder;

Lane 1, a 116-bp region of RYR1;

Lane 2, a 162-bp region of ACTA;

Lane 3, a 166-bp region of RSP18;

Lane 4, a 166-bp region of ENO3;

Lane 5, a 226-bp region of GAPDH;

Lane 6, a 232-bp region of OAZ1;

Lane 7, a 307-bp region of ACTB;

Lane 8, a 308-bp region of TNF.


5ml (50 reactions) QFR82805

50ml (500 reactions) QFR82050

QuickExtract FFPE RNA Extraction Solution, DNase buffer 1, RNase-Free DNase I, Stop Solution.

QuickExtractTM FFPE RNA Extraction kit

QuickExtractTM FFPE RNA Extraction kit

Tabel.1 QuickExtract FFPE RNA

QuickExtractTM FFPE RNA Extraction kit RNA extraction kit from FFPE sample

FFPE sample xylene paraffin RNA degradation

QuickExtractTM FFPE RNA Extraction kit paraffin cross

-linking 35

From FFPE Sample/ P.21P.21

QuickExtract RNA Extraction Kit

1 total RNA

phenolchloroform

DNase I

Fig.1 105 HeLa cell 100ul Quickextract RNA solution vortex 1 RNA 1ul RNA

lysate MMLV cDNA 2ul cDNA template p532 6 primer sets

PCR

Lanes M: 100-bp ladder

Lane 1: 12,984-13,892

Lane 2: 9,406-10,202

Lane 3: 5,194-5,802

Lane 4: 4,191-4,690

Lane 5: 2,280-2,676

Lane 6: 1,029-1,329

Fig.2 RNA extraction kit

RNA MMLV cDNA ALDOA primer set PCR

Lane M, 100 bp ladder ;

Lanes 1 and 2, QuickExtract RNA Extraction Kit;

Lanes 3 and 4, kit from Competitor 1;

Lanes 5 and 6, kit from Competitor 2.


5ml (50 reactions) QER09015

50ml (500 reactions) QER090150

QuickExtract RNA Extraction Solution, DNase buffer 1, RNase-Free DNase I, Stop Solution, RiboGuardTM

RNase Inhibitor.

QuickExtractTM RNA Extraction kit

QuickExtractTM RNA Extraction kit RNA 103~105 cells 100ul Quickextract

RNA solution vortex 1 RT-PCR

P.22P.22

NucleoTrap mRNA Mini/Midi

NucleoSpin mRNA Mini NucleoSpin mRNA Midi

Technology Affinity Chromatography Affinity Chromatography

Format Oligo (dT) latex-beads suspension Oligo (dT) latex-beads suspension

Sample material 250 g Total RNA 1000 g Total RNA

Fragment Size 50b - 20Kb 50b - 20Kb

Typical yeild 10 g mRNA 40 g mRNA

A260/280 1.9 - 2.1 1.9 - 2.1

Elution Volume 10 - 20 l 10 - 20l

Preparation Time 30min / 6 preps 30min / 6 preps

Binding Capacity 5 g poly(A) mRNA/mg beads 5 g poly(A) mRNA/mg beads

Product Information


NucleoSpin mRNA Mini 12 reactions 740655

NucleoSpin mRNA Mini 12 reactions 740656

oligo (dT) latex-beads mRNA

cultured cell mRNA RNA

30 gDNA mRNA

mRNA isolationRT-PCRin vitro transcription

mRNA isolation

MN NucleoTrap mRNA Kit Q S

mRNA

500ug total RNA from mouse kidney, Lane 1 & 2

Q mRNA kitLane 3 & 4NucleoTrap mRNA Kit

Lane 5 & 6 S mRNA KitLane 7 & 8

mRNA 2.5ug RNA 1.5% Formaldehyde agarose

gel 4.5-kb parkin gene transcript

(Parkinsons Disease) 32P-Labeled DNA

probe Northern Blot NucleoTrap

mRNA kit mRNA Q S

mRNA Isolation P.23P.23

mRNA-ONLY Prokaryotic/Eukaryotic mRNA Isolation Kit mRNA-ONLY Prokaryotic mRNA Isolation Kit with poly(A)-Tailing

mRNA rRNA RNA amplification

poly(A)-Tailing kit mRNA Poly(A)

mRNA-ONLY Prokaryotic/Eukaryotic isolation Kit Terminator 5-Phosphate-Dependent

Exonuclease total RNA rRNA tRNATerminator 5-Phosphate-Dependent Exonuclease

5 3exonuclase 5-monophosphat RNA Total RNA rRNA 5

-monophosphat Terminator 5-Phosphate-Dependent Exonuclease 1

mRNA mRNA RT-PCRgene expression analysisNorthern blot

RNA

mRNA-ONLY Prokaryotic isolation Kit with poly(A)-Tailing Terminator 5-Phosphate-

Dependent Exonuclease Poly(A) polymerase mRNA 3 ATP A-tailing

reaction cDNA gene expression analysis


mRNA-ONLY Prokaryotic mRNA Isolation Kit with poly(A)-Tailing 10 reactions MOT60510

mRNA-ONLY Prokaryotic isolation Kit 10 purifications MOP51010

24 purifications MOP51024

mRNA-ONLY Eukaryotic isolation Kit 10 purifications MOE51010

24 purifications MOE51024

: Terminator 5-Phosphate-Dependent Exonuclease, mRNA-ONLY Prokaryotic 10X Reaction Buffer, Ri-

boGuard RNase Inhibitor, mRNA-ONLY Stop Solution, 5 M LiCl Solution, RNase-Free Water.

E.coli total RNA(Lane1) 718nt RNA

mRNA-ONLY Prokaryotic/Eukaryotic isolation Kit

Terminator 5-Phosphate-Dependent Exonuclease

(Lane3) poly(A)-tailing mRNA (Lane4)

Lane5 total RNA and mRNA

Lane6 RNA mix poly(A)-tainking

Lane3 Lane5 Terminator 5-Phosphate-

Dependent Exonuclease total RNA

rRNA

mRNA Isolation P.24P.24

NucleoSpin RNA Clean-up/Clean-up XS

single reagent RNA

sample RT-PCR inhibitors

in vitro transcription NTP

RNA label reaction

RNA clean-up kit

RT-PCRDNA/RNA-based chip hybridizationin vitro transcriptionenzyme or

chemical labeling reaction

XS column 5ul RNA

Fig.1 NucleoSpin RNA Clean-up XS

100%

Lane 1: Crude RNA (100ul, 85ng/ul)

Lane 3: RNA Clean-up XS (20ul, 404ng/ul)

Lane 1 crude RNA NucleoSpin RNA

Clean-up XS major

band

>95% (Crude RNA 8500ng / Clean-up

RNA 8080ng)

B C

Fig.2 rat kidney total RNA amino-allyl-aRNA NucleoSpin RNA

Clean-up Q (A) Agilent Bioanalyzer

RNA NucleoSpin RNA Clean-up Q RNA

Q (A) NucleoSpin RNA Clean-up aRNA

array analysisB C

A MN

Q

RNA Ladder

RNA Clean-Up P.25P.25

RNA Clean-up

NucleoSpin RNA Clean-up NucleoSpin RNA Clean-up XS

Technology Silica membrane technology Silica membrane technology

Format Mini spin columns XS spin columns

Sample material Up to 200l Up to 300l

Fragment Size >200b >200b

Typical Recover 85 ~ 95% 85 ~ 95%

A260/280 1.9 - 2.1 1.9 - 2.1

Elution Volume 40 - 100 l 5 - 20l

Preparation Time 20min / 6 preps 20min / 6 preps

Binding Capacity 200 g 90 g

Product Information


NucleoSpin RNA Clean-up 10 reactions 740948.10


250 reactions 740948.250

NucleoSpin RNA Clean-up XS 10 reactions 740903.10


250 reactions 740903.250

RNA Clean-Up P.26P.26

Fig.3 MN NucleoSpin RNA Clean-up

RNA NucleoSpin RNA Clean-up XS

MN NucleoSpin RNA Clean-up XS Q

3 (51x : 16x) NucleoSpin RNA Clean-

up RNA column 4.5x RNA

NucleoSpin TriPrep column RNADNAProtein

RNA DNA

Fig.1 NucleoSpin Triprep DNA

A260/280 1.81~1.94

1. Lamda DNA marker HindIII 2. HeLa

3. Mouse Liver 4. Zebrafish 5. garden cress root

6. garden cress leave 7.Yeast 8. Bacteria

Fig.2 NucleoSpin Triprep RNA Agilent

2100 RNAHeLa cell RIN=9.5mouse liver

RIN=9.1

1. Lamda DNA marker HindIII 2. HeLa

3. Mouse Liver 4. Zebrafish 5. garden cress root

6. garden cress leave 7.Bacteria 8. Yeast

Fig.3 NucleoSpin Triprep protein SDS-PAGE

1. Lamda DNA marker HindIII 2. HeLa 3. Mouse Liver

4. Zebrafish 5. garden cress root 6. garden cress leave

7.Yeast 8. Bacteria

P.27P.27

NucleoSpin Triprep 10 / 50 / 250 reactions 740966.10/.50/.250


NucleoSpin RNA/Protein cultured cell RNA

DNase I RNA

RNA

gene expression analysissiRNA analysis

Fig.2 NucleoSpin RNA/Protein 106 HeLa cells

total RNA Agilent 2100 RNA RNA

quality

NucleoSpin RNA/Protein

Fig1. Inducible Knock-down of Hint1 in MCF-7 Tet-On cells

shRNA Hint1 silence Hint1 p53

A) 3 stable clonedempty vector (mock)vector with func-

tional shRNA (shRNA-Hint1 430/431) vector with inactive

shRNA (shRNA-Hint1 432/433) 1ug/ml dioxycycline

induction48 protein 50ug anti-Hint1

(anti-21) anti-b-actin (control) western bloting

B) & C) NucleoSpin RNA/Protein RNA

(B ) western bloting P53 b-actin

(control)RNA p53 primer b-actin primer (control)

RT-PCR C


NucleoSpin RNA /Protein 10/50/250 reactions 740933.10/.50/.250

P.28P.28

NucleoSpin RNA/DNA buffer set NucleoSpin RNA IINuclsoSpin RNA XSNucleoSpin RNA Plant NucleoSpin RNA/

Protein kit

sample RNA DNA

PCRRT-PCRreal-time PCR

NucleoSpin RNA/DNA buffer set

DNA RNA HeLa cells

NucleoSpin RNA II NucleoSpin RNA/

DNA buffer set 5x106 HeLa cells DNA

RNA denature gel(A,C) TAE agarose

gel (B,D)

Table.1 NucleoSpin RNA/DNA buffer set

DNA RNA A260/280 ratio

ARF1 primer set PCRRT-

PCR NucleoSpin RNA/DNA buffer set

HeLa cell DNA RNA ARF1 primer

set gDNA intron gDNA PCR

RNA RT-PCR product 207bp

+ : positive control - : negative control

P.29P.29

NucleoSpin RNA /DNA buffer set 100 reactions 740944


Bio-Genesis Technology Inc.

http://www.biogenesis.com.tw

[email protected] ;[email protected]

0800-211-667

02-26558877 037-687493

04-22602466 07-3422370 03-8463953

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2010創世紀專刊_RNA Extraction_100708

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